Biosystem dynamics and analysis of Rhodobacter sphaeroides strains with increased lipid accumulation
EMSL Project ID
48815
Abstract
We are requesting access to EMSL microscopy, including ETEM and STEM electron microscopes and the helium ion microscope, as well as the super resolution structured illumination fluorescence and multi-photon/FLIM integrated confocal microscopes. We also are requesting access to high-resolution proteomics. We propose to utilize these facilities, in collaboration with EMSL experts, to obtain molecular and high-resolution insights into overproduction and secretion of potential advanced biofuel precursors in bacteria. Specifically, we will characterize the location, organization and composition of accumulated intra-and extracellular lipids in “high lipid” producing mutants that we have isolated in the photosynthetic bacterium Rhodobacter sphaeroides. We are studying lipid production in R. sphaeroides since it can naturally increase fatty acid content under anaerobic conditions. Access to the state of the art imaging and proteomic facilities at EMSL is crucial to unraveling the biological processes leading to lipid accumulation in these strains. Our strategy is to use the insight we gain of fatty acid accumulation in R. sphaeroides to develop approaches to improve the yield of fatty acids or other fuel precursors in this or possibly other microbes. This project is supported, in part, by the Office of Biological and Environmental Research through its funding of Great Lakes Bioenergy Research Center (GLBRC) with the goal of generating knowledge needed to produce next generation biofuels or fuel precursors. EMSL supports this mission by providing innovative facilities and human resources to improve energy production. The proposed collaborative project leverages genome-enabled and system biology activities within GLBRC and the cutting-edge imaging and proteomic capabilities of EMSL to provide knowledge about how biofuel-producing bacteria store lipids. Data generated in a currently funded collaborative project demonstrate that there are striking differences in the ultrastructure and organization of lipids in some of our mutants that we are able to visualize using the unique capabilities at EMSL. Access to the EMSL state of the art microscopy facilities is crucial to the success of these studies, as GLBRC has neither the high-resolution facilities nor the expertise to conduct these studies. Use of the EMSL microscopy imaging facilities and high-resolution proteomics will allow us to gain critical insight into the dynamic processes leading to increased fatty acid content in our "high lipid" mutants of R. sphaeroides. This knowledge will be integral to ongoing studies that seek to elucidate the genetic and biochemical mechanisms that regulate the flow of strategic precursors into fatty acids in bacteria. These studies will help to improve strategies for modifying and manipulating microbes for fatty acid-derived biofuels and other bio-products of interest to DOE.
Project Details
Project type
Large-Scale EMSL Research
Start Date
2015-10-01
End Date
2017-09-30
Status
Closed
Released Data Link
Team
Principal Investigator
Team Members
Related Publications
Lemmer KC, A Dohnalkova, DR Noguera, and TJ Donohue. 2015. "Oxygen-??dependent Regulation Of Bacterial Lipid Production." Journal of Bacteriology 197(9):1649-1658. doi:10.1128/JB.02510-14
Lemmer KC, W Zhang, SJ Langer, A Dohnalkova, D Hu, RA Lemke, JS Piotrowski, G Orr, DR Noguera, and TJ Donohue. 2017. "Mutations that alter the bacterial cell envelope increase lipid production." mBio 8(3):Article No. e00513-17. doi:10.1128/mBio.00513-17